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Molecular Characterization of the Incompatible Interaction of Vitis Vinifera Leaves With Pseudomonas Syringae Pv. Pisi: Expression Of Genes Coding For Stilbene Synthase And Class 10 PR Protein

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Abstract

The interaction between Vitis vinifera and Pseudomonas syringae pv. pisi was examined at the pathological and molecular levels. Leaves infiltrated with the bacterial suspension developed necrotic regions which remained restricted to the infiltrated areas. In the infiltrated zone the number of bacteria decreased around 24h after inoculation whilst no bacteria could be isolated from the non-infiltrated zone. At the molecular level, two genes, stilbene synthase (SS) and a PR10 gene, encoding putative defense proteins, were analyzed. Expression of the SS gene, measured by the analysis of transcript accumulation, was shown to be highly induced and was followed by the accumulation of resveratrol (peaking at approximately 48h post-inoculation), considered as one of the major nobreak phytolaexins in the Vitis species. We report for the first time the isolation of a genomic clone (VvPR10-1) coding for a PR10 protein from this plant. The accumulation of the corresponding mRNA (0.8kb) was observed from 3 to 96h post-inoculation, peaking at 24–48h, and was followed by the accumulation (between 24 and, at least, 96h after inoculation) of the encoded polypeptide as detected by immuno-blotting. These results indicate that our experimental system based on an interaction of the non-host plant V. vinifera leaves with P. s. pv. pisi, has to be considered as an HR-like response and is well suited for the analysis of the defense reaction of this economically important species.

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References

  • Bavaresco L, Petegolli D, Cantu E, Fregoni M, Chiusa G and Trevisan M (1997) Elicitation and accumulation of stilbene phytoalexin in grapevine berries infected by Botrytis cinerea. Vitis 36: 77-83

    Google Scholar 

  • Bavaresco L, Fregoni C, Cantu E and Trevisan M (1999) Stilbene compounds: from the grapevine to wine. Drugs Exptl Clin Res XXV: 57-63

    Google Scholar 

  • Breda C, Sallaud C, El-Turk J, Buffard D, de Kozak I, Esnault R and Kondorosi A (1996) Defense reaction in Medicago sativa: a gene encoding a class 10 PR protein is expressed in vascular bundles. Molecular Plant-Microbe Interactions 9: 713-719

    Google Scholar 

  • Busam G, Kassemyer H-H and Matern U (1997) Differential expression of chitinases in Vitis vinifera L. responding to systemic acquired resistance activators or fungal challenge. Plant Physiology 115: 1029-1038

    Google Scholar 

  • Calderon AA, Zapata JM, Munoz R, Pedreno MA and Barcelo AR (1993) Resveratrol production as a part of the hypersensitivelike response of grapevine cells to an elicitor from Trichoderma viride. New Phytology 124: 455-463

    Google Scholar 

  • Chang M-M, Horovitz D, Culley D and Hadwiger LA (1995) Molecular cloning and characterization of pea chitinase gene expressed in response to wounding, fungal infection and the elicitor chitosan. Plant Mol Biol 28: 105-111

    Google Scholar 

  • Coutos-Thévenot P, Goebel-Tourand I, Mauro MC, Jouanneau JP, Boulay M, Deloire A and Guern J (1992) Somatic embryogenesis from grapevine cells. I - Improvement of embryo developments by changes in culture conditions. Plant Cell Tissue Organ Cult 29: 125-133

    Google Scholar 

  • Daniels CH, Fristensky B, Wagoner W and Hadwiger KA (1987) Pea genes associated with non-host disease resistance to Fusarium are also active in race-specific disease resistance to Pseudomonas. Plant Mol Biol 8: 309-316

    Google Scholar 

  • Dercks W and Creasy LL (1989) The significance of stilbene phytoalexins in the Plasmopora-grapevine interaction. Physiol Mol Plant Pathol 34: 189-202

    Google Scholar 

  • Derkel JP, Legendre L, Audran JC, Haye B and Lambert B (1996) Chitinases of the grapevine (Vitis vinifera L.): five isoforms induced in leaves by salicylic acid are constitutively expressed in other tissues. Plant Sci 119: 31-37

    Google Scholar 

  • Derkel JP, Audran JC, Haye B, Lambert B and Legendre L (1998) Characterization, induction by wounding and salicylic acid, and activity against Botrytis cinerea of chitinases and β-1,3-glucanases of ripening grape berries. Physiol Plant 104: 56-64

    Google Scholar 

  • Derkel JP, Baillieul F, Manteau S, Audran JC, Haye B, Lambert B and Legendre L (1999) Differential induction of grapevine defenses by two strains of Botrytis cinerea. Phytopathology 89: 197-203

    Google Scholar 

  • Esnault R, Buffard D, Breda C, Sallaud C, ElTurk J and Kondorosi A (1993) Pathological and molecular characterizations of alfalfa interactions with compatible and incompatible bacteria, Xanthomonas campestris pv. alfalfae and Pseudomonas syringae pv. pisi. Mol Plant-Microbe Interact 6: 655-664

    Google Scholar 

  • Fleuriet A and Macheix JJ (1972) Séparation et dosage par chromatographie en phase gazeuse de l'acide chlorogénique et des catechines des fruits. J Chromatogr 74: 339-345

    Google Scholar 

  • Gajhede M, Osmark P, Poulsen FM, Ipsen H, Larsen JN, van Neerven RJJ, Schou C, Lowenstein H and Spangfort MD (1997) X-ray and NMR structure of Betv1, the origin of birch pollen allergy. Nature Struct Biol 3: 1040-1045

    Google Scholar 

  • Goodwin PH, Hsiang T and Erickson L (2000) A comparison of stilbene and chalcone synthases including a new stilbene synthase gene from Vitis riparia cv. Gloire de Montpellier. Plant Sciences 151: 1-8

    Google Scholar 

  • Hain R, Reif HJ, Krause E, Langebartels R, Kindl H, Vorman B, Wiese W, Schmelzer E, Schreier PH, Stocker RH and Stenzel K (1993) Disease resistance results from foreign phytoalexin expression in a novel plant. Nature 361: 153-156

    Google Scholar 

  • Jeandet P, Bessis R and Gautheron B (1991) The production of resveratrol (3,5,40-trihydroxystilbene) by grape berries in different development stages. Am J Enol Vitic 42: 41-46

    Google Scholar 

  • Jeandet P, Bessis R, Sbagghi M and Meunier P (1995) Production of the phytoalexin resveratrol by grapes as a response to Botrytis attack under natural conditions. J Phytopathol 143: 135-139

    Google Scholar 

  • Kraeva E, Tesniére C, Terrier N, Romieu C, Sauvage FX, Bierne J and Deloire A (1998) Transcription of a β-1,3-glucanase gene in grape berries in a late developmental period, or earlier after wounding treatments. Vitis 37: 107-111

    Google Scholar 

  • Langcake P and Pryce RJ (1976) The production of resveratrol by Vitis vinifera and other members of the Vitaceae as a response to infection or injury. Physiol Plant Pathol 9: 77-86

    Google Scholar 

  • Langcake P and Pryce RJ (1977) The production of resveratrol and the viniferins by grapevines in response to ultraviolet irradiation. Phytochem 16: 1193-1196

    Google Scholar 

  • Langcake P, Cornford CA and Pryce RJ P (1979) Identification of pterostilbene as a phytoalexin from Vitis vinifera leaves. Phytochem 18: 1025-1027

    Google Scholar 

  • Langcake P (1981) Disease resistance of Vitis spp and the production of resveratrol, "-viniferin, -viviferin and pterostilbene. Physiol Mol Plant Pathol 18: 213-226

    Google Scholar 

  • Leckband G and Lorz H (1998) Transformation and expression of a stilbene synthase gene of Vitis vinifera L. in barley and wheat for increased fungal resistance. Theor Appl Genet 96: 1004-1013

    Google Scholar 

  • Mansfield J (1990) Recognition and response in plante/fungus interactions. Recognition and Response in Plant-Virus Interactions. Fraser RSS. Berlin. Springer-Verlag. H41. 31-52.

    Google Scholar 

  • Mauro MC, Toutain S, Walter B, Pinck L, Otten L, Coutos-Thévenot P, Deloire A and Barbier P (1995) High efficiency regeneration of grapevine plants transformed with the GFLV coat protein gene. Plant Sci 112: 97-106

    Google Scholar 

  • Melchior F and Kindl H (1990) Grapevine stilbene synthase cDNA only slightly differing from chalcone synthase cDNA is expressed in Escherichia coli into a catalytically active enzyme. FEBS Lett 268: 17-20

    Google Scholar 

  • Melchior F and Kindl H (1991) Coordinate-and elicitordependent expression of stilbene synthase and phenylalanine ammonia-lyase genes in Vitis cv. Optima. Arch Biochem Biophys 288: 552-557

    Google Scholar 

  • Moiseyev GP, Fedoreyeva LI, Zhuravlev YN, Yasnetskaya E, Jekel PA and Beintema JJ (1997) Primary structures of two ribonucleases from ginseng calluses: New members of the PR-10 family of intracellular pathogenesis-related plant proteins. FEBS Lett 407: 207-210

    Google Scholar 

  • Ni W, Sewalt VJH, Korth KL, Blount JW, Balance GM and Dixon RA (1996) Stress response in alfalfa. XXI. Activation of caffeic acid 3-O-methyltransferase and caffeoyl coenzyme A 3-O-methyltransferase genes does not contribute to changes in metabolite accumulation in elicitor-treated cell-suspension cultures. Plant Physiol 112: 717-726

    Google Scholar 

  • Rasmussen JB, Hammerschmidt R and Zook MN (1991) Systemic induction of salicylic acid accumulation in cucumber after inoculation with Pseudomonas syringae pv. syringae. Plant Physiol 97: 1342-1347

    Google Scholar 

  • Renault A-S, Deloire A and Bierne J (1996) Pathogenesis-related proteins in grapevine induced by salicylic acid and Botrytis cinerea. Vitis 35: 49-52

    Google Scholar 

  • Risler JL, Delorme MO, Delavroix H and Hénaut A (1988) Amino acid substitutions in structurally related proteins as a pattern recognition approach. J Mol Biol 11: 1019-1029

    Google Scholar 

  • Robinson SP, Jacobs A K and Dry IB (1997) Aclass IV chitinase is highly expressed in grape berries during ripening. Plant Physiol 114: 771-778

    Google Scholar 

  • Sallaud C, El Turk J, Breda C, Buffard D, de Kozak I, Esnault R and Kondorosi A (1995) Differential expression of cDNA coding for chalcone reductase, a key enzyme of the 5-deoxyflavonoid pathway, under various stress conditions in Medicago sativa. Plant Sciences 109: 179-190

    Google Scholar 

  • Sallaud C, Zuanazzi J, El-Turk J, Leymarie J, Breda C, Buffard D, de Kozak I, Ratet P, Husson P, Kondorosi A and Esnbault R (1997) Gene expression is not systematically linked to phytoalexin production during alfalfa leaf interaction with pathogenic bacteria. Mol Plant-Microbe Interact 10: 257-267

    Google Scholar 

  • Salzman RA, Tikhonova I, Bordelon BP, Hasegawa PM and Bressan RA (1998) Coordinate accumulation of antifungal proteins and hexoses constitutes a developmentally controlled defense response during fruit ripening in grape. Plant Physiol 117: 465-472

    Google Scholar 

  • Sarig P, Zutkhi Y, Monjauze A, Lisker N and Ben-Arie R (1997) Phytoalexin elicitation in grape berries and their susceptibility to Rhizopus stolonifer. Physiol Mol Plant Pathol 50: 337-347

    Google Scholar 

  • Schröder G, Brown JWS and Schröder J (1988) Molecular analysis of resveratrol synthase: cDNA, genomic clones and relationship with chalcone synthase. Eur J Biochem 172: 161-169

    Google Scholar 

  • Schröder J and Schröder G (1990) Stilbene and chalcone synthase: related enzymes with key functions in plants-specific pathways. Z Naturforsch 45c: 1-8

    Google Scholar 

  • Schubert R, Fischer R, Hain R, Schreier PH, Bahnweg G, Ernst D and Sandermann H (1997) An ozone-responsive region of the grapevine resveratrol synthase promoter differs from the basal pathogen-responsive sequence. Plant Mol Biol 34: 417-426

    Google Scholar 

  • Sikorski MM (1997) Expression of Lupinus luteus cDNA coding for PR10 protein in Escherichia coli: purification of the recombinant protein for structural and functional studies. Acta Biochim Pol 44: 565-578

    Google Scholar 

  • Stark-Lorenzen P, Nelke B, Hänssler G, Mühlbach HP and Thomzik JE (1997) Transfer of grapevine stilbene synthase gene to rice (Oryza sativa L.). Plant Cell Reports 16: 668-673

    Google Scholar 

  • Tattersall DB, van Heeswijck R and Hoj PB (1997) Identification and characterization of a fruit-specific thaumatin-like protein that accumulates at very high level in conjunction with the onset of sugar accumulation and berry softening in grapes. Plant Physiol 114: 759-769

    Google Scholar 

  • Thomzik JE, Stenzel K, Schreier PH, Hain R and Stahl DJ (1997) Synthesis of grapevine phytoalexin in transgenic tomatoes (Lycopersicum esculentum Mill.) conditions resistance against Phytophthora infestans. Physiol Mol Plant Pathol 51: 265-278

    Google Scholar 

  • Van Loon LC, Pierpoint WS, Boller T and Conejero V(1994) Recommendations for naming plant pathogenenesis-related proteins. Plant Mol Biol Reporter 12: 245-264

    Google Scholar 

  • Wen J, Vanek-Krebitz M, Hoffmann-Sommerbruger K, Scheiner O and Breiteneder H (1997) The potential of Betv1 homologues, a nuclear multigene family, as phylogenetic markers in flowering plants. Mol Phylogen Evol 8: 317-333

    Google Scholar 

  • Wiese W, Vornam B, Krause E and Kindl H (1994) Structural organization and differential expression of three stilbene synthase genes located on a 13 kb grapevine DNA fragment. Plant Mol Biol 26: 667-677

    Google Scholar 

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Correspondence to Robert Esnault.

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Robert, N., Ferran, J., Breda, C. et al. Molecular Characterization of the Incompatible Interaction of Vitis Vinifera Leaves With Pseudomonas Syringae Pv. Pisi: Expression Of Genes Coding For Stilbene Synthase And Class 10 PR Protein. European Journal of Plant Pathology 107, 249–261 (2001). https://doi.org/10.1023/A:1011241001383

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